Collection/extraction container for biological material in forensic samples

ABSTRACT

Relates to a collection/extraction container ( 1 ) for collecting solid forensic samples ( 2 ) and/or for extracting biological material from these solid forensic samples ( 2 ) and its use. The container ( 1 ) has a top opening ( 3 ) for the insertion of a solid forensic sample ( 2 ). The container ( 1 ) comprises a basket ( 7 ) and an essentially horizontal intermediate floor ( 8 ) for the retention of the solid forensic sample ( 2 ) during digestion/lysis and extraction, the intermediate floor ( 8 ) being permeable to fluids and dividing the inner chamber ( 5 ) into an upper sample space ( 9 ) and a lower fluid space ( 10 ). The container according to the invention is characterized in that is comprises an essentially vertical channel ( 11 ), which is accomplished to allow a pipetting needle ( 12 ) to be inserted through the sample space ( 9 ) and into the fluid space ( 10 ) in that the channel ( 11 ) has a top orifice ( 13 ) that is situated nearby a top opening ( 3 ) of the container ( 1 ) and in that the channel ( 11 ) penetrates the intermediate floor ( 8 ) in a bottom orifice ( 14 ), wherein the container ( 1 ) also comprises a partition wall ( 15 ) that at least partially surrounds the channel ( 11 ) and that separates the latter from the upper sample space ( 9 ).

The present invention relates to a collection/extraction container forcollecting solid forensic samples and/or for extracting biologicalmaterial from these forensic samples. This container has a top openingfor the insertion of a forensic sample. The top opening is surrounded byan essentially vertical wall, which encloses an inner chamber that islimited in depth by a bottom. The wall and the bottom of the containerare impermeable to fluids. The container also comprises a basket and anessentially horizontal intermediate floor for the retention of the solidforensic sample during washing, i.e., digestion/lysis and extraction.The intermediate floor is permeable to fluids and divides the innerchamber into an upper sample space and a lower fluid space.

The collection of samples containing nucleic acid in order to determinethe genetic code is currently increasingly gaining in significance. Inconnection with combating crime, the collection of genetic samples inthe meaning of a “genetic fingerprint” is becoming more and moreimportant in two regards: firstly, the genetic code is to be acquiredfrom criminals who have already been arrested and secondly the acquireddata is to be compared to unknown traces found at a crime scene, forexample. In the first case, fresh and clearly identified samples areused to build up a data bank, which may be accessed in the second case.However in the second case, the collected samples are often incomplete,contaminated, and damaged, thus complicating the forensic work.Additionally, individual countries have already begun to collectbiological samples in order to preventively acquire the genetic code ofall individuals entering the country or even all the inhabitants of thecountry.

The attribute “forensic” refers to anything, which has a legal orcriminological character. The term is thus not only restricted to thefields of criminal law (e.g., legal medicine), but rather comprises anyprofessional activity within any legal proceeding. Forensically relevantsamples also comprise proteins (e.g., the prions causingCreutzfeld-Jacob syndrome, or bovine spongiform encephalopathy, or BSErespectively), viruses, bacteria, and other microorganisms, human oranimal bodily fluids (such as blood, sputum, feces, sperm, and urine),and single cells (such as oral mucosa cells and hair follicles).

Methods for isolating and analyzing human deoxyribonucleic acid (DNA)ribonucleic acids (RNA) are known from the prior art (cf., for example,Molecular Diagnostics: Isolation and Analysis of Human Genomic DNA, 1998Promega Notes No. 68, p. 20). These methods comprise the PCR methods(PCR=Polymerase Chain Reaction) well known per se for increasing thesample yield and thus the sensitivity of the analysis.

Containers for collecting samples and for manually preparing thecollected samples for an analysis or a PCR reaction are known from WO2004/105949 A1. Containers for performing the collection and subsequentPCR reaction in the field are known from US 2004/0214200 A1. However,these known methods appear quite complicated and the containers appearunsuitable for automated and/or robotic processing of samples.

Automated nucleic acid isolation is e.g., known from U.S. Pat. No.5,863,801. However, the device described in this patent is designed tobe used as a stand-alone device and does not work with the existingautomation and instruments typically used in forensics and researchlaboratories, such as a TECAN automated liquid handler (TECAN Schweiz AG, Seestrasse 103, CH-8708 Mannedorf, Switzerland). Given the resourcesrequired for such a laboratory to validate their processes, it is ofsignificant value to have a consumable that can closely mimic theirvalidated manual processes and be used on existing instrumentation.

A device for incubation, centrifugation and separation of DNA samples ona solid support is known as the Slicprep™ 96 device from PROM EGA Corp.(2800 Woods Hollow Road, Madison, Wis. 53711 USA). This device is madefrom polypropylene and is based on a 96-deep well microplate, a 96 spinbasket unit, and a collar for raising the baskets. In the incubationmode, the solid supports or forensic samples (e.g. dried buccal swabs)are placed in the baskets that are fully inserted in the deep wellplate. Digestion buffer or lysis buffer is added to cover the samplesand the sealed unit is incubated at a desired temperature. The basketsare then raised and a collar is inserted so that centrifugation in aswinging plate rotor will remove the extract, i.e., the DNA-containingsolution from the solid supports or forensic samples. After removal ofthe 96-spin basket unit and the collar, the deep well plate can then beplaced on a workstation for purifying the DNA. Although this device hasbeen designed to make DNA extraction from forensic samples moreautomation compatible, manual procedure steps are still required foradding the collar and then later removing the collar and the basketunit.

Nucleic acid isolation that is carried out similarly as just describedand that also requires significant manual steps during the process isknown from FITZCO Inc. (4300 Shoreline Drive, Spring Park, Minn. 55384,USA). The FITZCO Spin-eze™ device is a single tube device consisting ofa basket unit and an Eppendorf type receiving tube. In the initialextractions steps, solid supports containing DNA (i.e., swabs) areplaced in the basket, which is then inserted into the Eppendorf typetube. Digestion or lysis buffers are added to cover the forensic samplesand the sealed unit is incubated and later centrifuged to make theDNA-containing solution flow from the basket to the tube. The basket isthen manually removed and the tube can be placed on a workstation forpurifying the DNA.

Another system that is known from TECAN consists of two tubes, which canbe attached end-to-end with a “filter”, placed between them forprocessing. A sample tube contains the swab or other DNA containingforensic material. Buffers are added to the sample tube for initialprocessing and a receiving tube is attached upside down to the top ofthe sample tube. A rack with an array of such “doubletubes”is moved intoa “flipping” device, which rotates the two connected tubes so that theliquid can flow via the filter from the now upper sample tube into thelower receiving tube during a subsequent centrifugation. The tubes thenmay be separated. Also this system needs manual manipulation and inaddition, special devices for flipping the tubes are required.

An object of the present invention is to suggest an alternative deviceand/or an alternative method, which enable bound molecules, such asnucleic acids, to be processed and removed fully automatically fromsolid supports, such as buccal swabs.

This object is achieved according to a first aspect by acollection/extraction container for collecting solid forensic samplesand/or for extracting biological material from these forensic samplesaccording to the features of claim 1. The container has a top openingfor the insertion of a forensic sample. The top opening is surrounded byan essentially vertical wall, which encloses an inner chamber that islimited in depth by a bottom. The wall and the bottom are impermeable tofluids. The container comprises a basket and an essentially horizontalintermediate floor for the retention of the solid forensic sample duringwashing, i.e., digestion/lysis and extraction, the intermediate floorbeing permeable to fluids and dividing the inner chamber into an uppersample space and a lower fluid space.

The container according to the present invention is characterized inthat it comprises an essentially vertical channel, which is accomplishedto allow a pipetting needle to be inserted through the sample space andinto the fluid space in that the channel has a top orifice that issituated nearby the top opening of the container and in that the channelpenetrates the intermediate floor in a bottom orifice, wherein thecontainer also comprises a partition wall that at least partiallysurrounds the channel and that separates the latter from the uppersample space.

This object is achieved according to a second aspect according to thefeatures of claim 17, in that a method for extracting biologicalmaterial from forensic samples is suggested. The method according to thepresent invention comprises the following method steps:

-   (a) Providing at least one container 1 with an accessible top    opening 3, an upper sample space 9 with a basket 7, a fluid space    10, and a channel 11;-   (b) Placing solid forensic samples 2 through the top opening 3 into    the basket 7 in the upper sample space 9 of the at least one    container 1;-   (c) Adding digestion/lysis buffer into the basket 7 so that both the    lower fluid space 10 of the container 1 is filled and the upper    sample space 9 is also filled enough to cover the forensic sample 2;-   (d) Sealing the entire device 1 and then allowing to incubate to    enable the extraction process to occur;-   (e) Unsealing or opening the device 1 after incubation and removing    a volume of digestion buffer by accessing the bottom of the lower    fluid space 10 through the essentially vertical channel 11;-   (f) Placing the device 1 in a centrifuge and spinning so that any    remaining buffer in the solid forensic sample 2 collects down in the    bottom of the inner chamber 5;-   (g) Removing the remaining volume of digestion buffer that has    collected at the bottom of the lower fluid space 10 by the    centrifugation by access through the essentially vertical channel    11,    wherein the solid forensic samples 2 are retained in the basket 7 in    the sample space 9 of the at least one container 1 during all the    steps (c) through (g).

The volume removed preferably is combined with the volume removedearlier. This combined sample is now ready for purification of nucleicacids using commercial kits.

Additional preferred features according to the present invention resultfrom the dependent claims.

Advantages of the present invention comprise:

-   -   The same container that is used for extraction of biological        material from e.g., the surfaces of solid forensic samples can        also be used for the collection of said forensic samples in the        field.    -   The container provides a port or channel that enables access to        the bottom of the container for a pipette tip without requiring        removal of the basket that contains the solid forensic sample.    -   The container can be racked into an array of e.g., 6×8 tubes and        utilized fully automated, as there is no need to assemble or        disassemble parts during processing. Thus, common automation        devices such as centrifuges, robotic arms, PCR thermostatic        tables etc. can be used.    -   There is no need for special devices for flipping the tubes or        racks that contain the tubes.    -   Labeling the container with an identification tag provides clear        identification of a particular forensic sample and the        biological material extracted from this solid forensic sample.

The present invention will be explained in greater detail on the basisof exemplary embodiments and schematic drawings, which do not restrictthe scope of the present invention. Whereas it is shown in:

FIG. 1 vertical partial sections through two different racks having fourdifferent embodiments of the container according to the presentinvention:

FIG. 1A shows flat bottom containers with friction fit inserted baskets;

FIG. 1B shows a container shaped as a PCR tube and a containerconsisting from two parts;

FIG. 2 horizontal partial sections through the two racks of FIG. 1:

FIG. 2A shows a collecting rack with friction fit inserted containers;

FIG. 2B shows an extracting rack with containers held in place byretention means;

FIG. 3 vertical partial sections through a rack having two differentembodiments of the container according to the present invention.

FIG. 1 shows vertical partial sections through two different rackshaving four different embodiments of the container 1 according to thepresent invention. FIG. 1A exhibits flat bottom containers with frictionfit inserted baskets. Both containers shown are collection/extractioncontainers 1 for collecting solid forensic samples 2 and/or forextracting biological material from these forensic samples 2. Solidforensic samples 2 are typically selected from a group, which comprisesa gauze pad or buccal swab; a finding; a filter paper or WHATMAN FTApaper (Whatman plc., 27 Great West Road, Brentford, Middlesex, TW8 9BW,UK) respectively; a solid, sheet-like material for adsorbing nucleicacids or other biological molecules, such as preparation gels andcellulose membranes; and a textile piece. In most of the cases thebiological material sample to be extracted is selected from a groupcomprising bodily fluids, cells, DNA, RNA, proteins, microorganisms, andviruses. It is especially preferred however, to collect nucleic acidmaterial such as DNA and/or RNA with a buccal swab and to extract thenucleic acid material from this buccal swab within the container 1 ofthe invention.

Each container 1 has a top opening 3 for the insertion of a solidforensic sample 2, the top opening 3 being surrounded by an essentiallyvertical wall 4, which encloses an inner chamber 5 that is limited indepth by a bottom 6. The wall 4 and the bottom 6 are impermeable tofluids. Each container 1 comprises a basket 7 and an essentiallyhorizontal intermediate floor 8 for the retention of the solid forensicsample 2 during washing, i.e., digestion/lysis and extraction. Theintermediate floor 8 is permeable to fluids and divides the innerchamber 5 into an upper sample space 9 and a lower fluid space 10.

“Fluids” in the context of the present invention are to be understood asliquids, gases, or liquid/gas mixtures. “Solid” is interpreted as hardor soft material other than fluidic. A “sample” preferably is a solidforensic sample, but it can be any other sample that contains biologicalmaterial. A “finding” can be an artifact or casework sample as found ata criminal scene or any other non-specific solid material that containsbiological material that can be extracted.

The container 1 according to the invention comprises an essentiallyvertical channel 11, which is accomplished to allow a pipetting needle12 to be inserted through the sample space 9 and into the fluid space10. This is accomplished in that the channel 11 has a top orifice 13that is situated nearby the top opening 3 of the container 1 and in thatthe channel 11 penetrates the intermediate floor 8 in a bottom orifice14. The container 1 also comprises a partition wall 15 that at leastpartially surrounds the channel 11 and that separates the latter fromthe upper sample space 9.

Preferably, the wall 4 of the container 1 is essentially cylindrical atleast in the region of the upper sample space 9. It is also preferredthat the channel 11 is arranged off-center and partially formed by thewall 4 and the partition wall 15 in order to leave as much space aspossible for a solid forensic sample 2 that can be introduced into theupper sample space 9 and that can be kept here by the basket 7. It isespecially preferred that the basket 7 of the container 1 is sized toaccommodate a buccal swab.

As the channel 11 preferably is arranged off-center, it defines anessentially vertical off-center axis 40. The bottom 6 and/or the lowersection 17 of the wall 4 thus are preferably accomplished to form afluid collecting area 21, which is at a lower level then the rest of thebottom 6 of the container 1, the fluid collecting area 21 lying on thisessentially vertical off-center axis 40.

As shown in FIG. 1A, the basket 7 may be accomplished as a singleinjection molded piece of polymer material that is inserted into theinner chamber 5 of the container 1. Alternately as shown in FIG. 1B (seethere on the right side), the basket 7 is accomplished as a part of anupper section 16 of the wall 4 and formed together with this uppersection 16 of the wall 4 as a single injection molded piece of polymermaterial. The lower section 17 of the wall 4 advantageously comprises atits upper end 18 a socket 19 into which a lower end 20 of the uppersection 16 of the wall 4 can be sealingly inserted. As also depicted inFIG. 1B (see there on the left side), at least in the region of thelower fluid space 10, a container 1 may be shaped as a PCR tube.

It is preferred that the bottom 6 of the container 1 is accomplished toform together with the wall 4 or a lower section 17 of the wall 4 asingle injection molded piece of polymer material. The preferred polymermaterial for injection molding of the parts of the container 1 accordingto the invention is polypropylene. Alternately, the bottom 6 of thecontainer 1 is accomplished as a film structure 27 that is glued orwelded to the wall 4 or to a lower section 17 of the wall 4 (see e.g.,FIG. 1A, left side).

The bottom 6 preferably comprises a flat area 22 on its outer side, onor in which flat area 22 an information tag 23 is located (see e.g.,FIG. 1B, right side). The information tag 23 advantageously is selectedfrom a group comprising a 1-D bar code, a 2-D bar code, an RFIDtransponder, and a RuBee transceiver. The principle of 1-D and 2-Dbarcodes is well known to those skilled in the art and is based on theoptical scanning of a high-contrast identification marking. Theadvantage of such identification is the relatively simple physicalprinciple; however, there must be a visual contact between the scanningdevice and the information tag.

RFID transponders are also known per se and operate at high frequency(HF, such as 900 MHz) or ultra-high frequency (UHF). They transmit andreceive radio signals, while the newer RuBee transceivers operate atwavelengths below 450 kHz and emit and receive signals, which areprimarily based on magnetism. The passive RFID transponders may receiveapproximately 100 (HF) or 150-200 (UHF) messages per second. Incontrast, the active RuBee transceivers may only receive approximately10 messages/second; visual contact is not needed in any case. The typeof information tag 23 used is thus a function, inter alia, of thedensity of the data transfer and the presence of a visual contact. Inthe case of FIG. 1B, an RFID transponder or a RuBee transceiver issituated on the vertical wall 4 of the container 1 (see left side),while a 2-D barcode is situated on the flat area 22 of the bottom 6 (seeright side). Any arbitrary combination of bar codes and RFID transponderor a RuBee transceiver are conceivable, this is especially the case whene.g., a simple 1-D bar code is used to identify a forensic sample duringcollection in the field and an RFID transponder or a RuBee transceiveris utilized on the same tube during automatic extraction of the relevantbiological material in a high through put workstation of a forensiclaboratory.

The container 1 preferably comprises a closure 24 for the top opening 3.Advantageously depending from the site of use, this closure 24 isselected from a group, which comprises a stopper 25, a cover 26, and afilm 27. As an example, a collecting set 28 is shown in FIG. 1A. Thiscollecting set 28 has at least one collection/extraction container 1according to the invention. This collecting set 28 comprises acollecting rack 29 having compartments 30 for receiving the containers.

Preferably, the compartments 30 of this collecting rack 29 are open onthe top and bottom, so that the containers 1 are insertable from aboveor below into a compartment 30 and are ejectable downward or upward fromthis compartment. It is especially preferred that the collecting rack 29has the dimensions of a standard microplate. Standards for thedimensions of microplates have formally been published from ANSI(American National Standards Institute). These standards define amicroplate according to the Footprint Dimensions (ANSI/SBS 1-2004), theHeight Dimensions (ANSI/SBS 2-2004), the Bottom Outside FlangeDimensions (ANSI/SBS 3-2004), and the Well Positions (ANSI/SBS 4-2004).However, strip racks with six or eight compartments in one row may alsobe chosen for collecting forensic samples.

If the collecting set 28 originally contains sample tubes, or containers1 respectively, without individual closure, a film 27 that covers theentire collecting rack 29 on the bottom as well as on the top ispreferred (see FIG. 1A, left). Thus, perfectly clean containers that arefree from any contamination are provided at the site of forensic samplecollection. As soon as a fresh sample 2 has been introduced into thebasket 7 of a container 1, a stopper 25 may be mounted to entirely closethe top opening 3 of the container 1 (see FIG. 1A, right). The stopper25 may contain a bar code (not shown here) that identifies theindividual forensic sample 2 in the container 1.

As an alternative, the collecting set 28 may originally contain sampletubes or containers 1 with individual closure 25. In this case, at leastthe film 27 at the top of the collecting rack 29 can be dispensed with.If also the bottom of the compartments 30 is not covered by a film 27,the bar code can be supplied on the flat area 22 of the container bottom6 (not shown here). Eventually, no bar code at all can be used in cases,where the containers are labeled with only an RFID transponder or aRuBee transceiver. Such intelligent labels can also contain additionaldata that are entered prior to or during the sample collection and allfurther processing and that are readable with a dedicated device.

In the laboratory, the samples can be treated in the collecting rack 29.It is however preferred, to concentrate the containers from differentcollecting racks 29 and to enter them into one or more extracting racks33. Such extracting racks 33 preferably have the shape and size of astandard microplate in order to enable transportation of theseextracting racks 33 by microplate robots and handling these extractingracks 33 in microplate handling stations of laboratory workstations. Anextracting rack 33 and at least one container 1 inserted into acompartment 30 of this rack forms an extracting set 32.

The containers 1 having the solid forensic sample 2 provided therein maybe inserted into arbitrary compartments 30 of such extracting racks 33by a robot (not shown). In order to accomplish this, a collecting rack29 and an extracting rack 33 are placed on top of each other such thatthe compartments 30 of both racks 29,33 are in register with each other.Depending now which one of the two racks is on top of the other; a robottool is used to push the containers 1 into the compartments of theextracting rack 33 from above of from below. This action is greatlyfacilitated in that both types of racks 29,33 have compartments 30 thatare also open on the top and the bottom, so that the containers 1 areinsertable from the bottom or top into a compartment 30 and areejectable downward or upward from this compartment 30.

Even when processing of the containers 1 is preferred to be carried outin racks 29,33, processing each container 1 individually by a robot ispossible too. That is, the containers 1 not necessarily have to beracked to be processed in an automated fashion. Actually, TECANinstruments, for instance, can move individual tubes using a so-called“Pick & Place Arm”. In deed, for casework samples in forensicsinvestigations, it is preferable to treat smaller numbers (e.g., 1 to 8)of samples simultaneously rather then wait for 96 samples. Even withsmaller numbers of samples, automation provides advantages such ashigher reproducibility, “chain of custody”, and walk away time.

As seen from the horizontal partial sections through the two racks 29,33 in FIG. 2, the containers 1 are preferably retained in thecompartments 30 of the racks 29,33 using retention means 35 (see FIG.2B). However, the containers 1 may sit in the compartments by frictionfit only (see FIG. 2A). The containers 1 preferably have an upper flange36 and a lower thickened area 37 that advantageously have identicalexternal diameters. This diameter is slightly larger than the distancebetween two parallel intermediate walls 34 of the collecting orextracting rack 29,33. Thus, the insertion of a container slightlydeforms this container and the four adjacent intermediate walls 34 andthe friction between the surfaces of this upper flange 36 and lowerthickened area 37 and the inner surfaces of the intermediate walls 34keeps the container inside a compartment 30.

The FIGS. 2A and 2B are horizontal partial sections through the tworacks 29,33 as shown in FIG. 1. In the right compartment of FIG. 1A andin the left compartment of FIG. 1B, a container 1 is inserted which'sbottom 6 (see FIG. 1A) or which's lower section 17 of the wall 4 (seeFIG. 1B) are accomplished to form a fluid collecting area 21, which isat a lower level then the rest of the bottom 6 of the container 1. Thisfluid collecting area 21 lies on the essentially vertical off-centeraxis 40. Whereas the relative position of the two other containers withrespect to the inserted basket 7 (see FIG. 1A, left) or with respect tothe integrated basket 7 (see FIG. 1B, right) is of no concern, therelative position of the channels 11 of the two containers with fluidcollecting areas 21 is important. In order to define a correct relativeposition of the basket 7, channel 11 and the respective collecting areas21 in the bottom 6 of the container 1, the basket preferably is equippedwith an essentially vertical orientation bar 41. The container containsin the region of the upper sample space 9 an essentially verticalorientation grove 42 that is shaped to accommodate the orientation bar41 of the basket 7. With this design of an orientation bar and grove41,42, the desired position of the basket 7 may be defined in itsorientation around the central axis 43 of the container 1 as well as inits height, i.e., its distance from the bottom 6 of the container 1.

Departing from the embodiment of an intermediate floor 8 as shown herewith square through-openings, but not departing from the spirit of thepresent invention, the intermediate floor can also be made permeable toliquids by through-openings that are of any other shape than square.Different material than the Polymer used for molding the basket 7 and/orthe container 1 can be used for the intermediate floor 8. Such materialcomprises filter paper and fabrics, which comprise natural and/orsynthetic fibers and/or metal wires. However, as the containers 1 aswell as the baskets 7 preferably are designed as low-cost consumables,the utilization of only one material for the production of the wholecontainer and basket is particularly preferred.

The desired position of the basket 7 with respect to its distance fromthe bottom 6 of the container 1 can also be defined in that a shoulder44 is implemented to the inner surface of the wall 4. The inner diameterof the upper sample space 9 is larger in this case than the innerdiameter of the lower fluid space 10 of the container 1, this formingthe shoulder 44. Accordingly, the outer diameter of the basket 7 islarger than the inner diameter of the lower fluid space 10, the basket 7thus resting on the shoulder 44 (see FIG. 1A, left).

FIG. 3 shows vertical partial sections through an extracting rack 33having two different embodiments of the container 1 according to thepresent invention. The direction of the section and viewing is indicatedin FIG. 2B with arrows. Also these containers 1 preferably have an upperflange 36 and a lower thickened area 37 with identical externaldiameters. This upper flange 36 and this lower thickened area 37 producea friction lock with retention means 35, which are preferably situatedin the corners of the compartments 30 and extend essentially over theentire height of the compartments 30. Thanks to this friction lock, thecontainers are securely retained in the compartments 30 and may not fallout spontaneously. In addition, these retention means 35 may have firstand second protrusions 38,39, between which the lower thickened area 37of the container 1 snaps upon insertion into the compartment 30, so thata specific height position of the containers 1 in the compartment 30 ispredefined and the retention in this defined position is reinforcedfurther. The friction lock with the containers 1 to be inserted may bedefined by the precise dimensions of these retention means 35, but alsoby the extent of their flexibility.

Whereas the container 1 in the left compartment of FIG. 3 is identicalwith the one in the right compartment of FIG. 1B, the position and sizeof the socket 19 at the upper end 18 of the lower section 17 of the wall4 are different. I.e., the sockets position is lower and the height ofthe socket 19 is larger. In both cases however, the intermediate floor 8is accomplished as a lattice of webs that integrally are produced in onepiece with the upper section 16 of the wall 4. The ability to deliver asealing between the two sections of the wall 4 increases with the sizeof the socket 19.

The container 1 preferably comprises a closure 24 for the top opening 3.Advantageously depending from the site of use, this closure 24 isselected from a group, which comprises a stopper 25, a cover 26, and afilm 27. As an example, a collecting set 28 is shown in FIG. 1A. Thiscollecting set 28 has at least one collection/extraction container 1according to the invention. This collecting set 28 comprises acollecting rack 29 having compartments 30 for receiving the containers.

In order to be able to automatically add a liquid to the solid sample, apipetting needle 12 can be introduced into the liquid space 10 via thechannel 11 (see FIG. 1B, left) or a dispenser needle can be positionedabove the solid forensic sample 2 (see FIG. 3). Also in order toautomatically withdraw a liquid from the liquid space 10, a pipettingneedle 12 can be introduced via the channel 11 (see FIG. 1B, left). Inall cases, access through the top opening 3 of the container 1 isnecessary. The top opening 3 may be entirely closed by a stopper 25 thathad been mounted after collecting the forensic sample (see FIG. 1B,right) or that was put onto the container in-between two preparationsteps in the lab workstation. Alternately, the top opening 3 may bypartly closed by a cover 26 that covers only the sample space 9 and thatleaves the channel 11 open (see FIGS. 1B and 2B, left). If the container1 contains in the region of the upper sample space 9 an essentiallyvertical orientation grove 42 that is shaped to accommodate theorientation bar 41 of the basket 7, the cover 26 preferably alsocomprises such an orientation bar 41 (see FIG. 2B, left).

All stoppers 25 and covers 26 preferably are equipped on their top witha grasping tube 31 (see e.g., FIG. 1B), which can be grasped by a robot(not shown) in order to remove the stoppers 25 or covers 26 or in orderto mount them appropriately in the containers 1. Departing from theembodiment shown but not from the spirit of the present invention, somesort of tab (e.g., like on the cover of an “Eppendorf” tube, not shown)that enables a person to easily open or remove a stopper 25 or cover 26can be utilized as an alternate solution. A user can then open thecontainers 1 manually before placing them on the automation system.Again, another embodiment may include a stopper 25 or cover 26 with apierceable septum above the basket 7 and/or channel 11 so that once thesample is placed in the basket, the stopper 25 or cover 26 does not needto be removed and the sample space 9 and/or the channel 11 can beaccessed by piercing the septum on the stopper 25 or cover 26 (notshown).

The present invention also provides a method for extracting biologicalmaterial from solid forensic samples 2, the method according to theinvention comprises:

-   (a) Providing at least one container 1 with an accessible top    opening 3, an upper sample space 9 with a basket 7, a fluid space    10, and a channel 11, according to the invention;-   (b) Placing solid forensic samples 2, typically a swab head, cloth,    WHATMAN FTA paper, or other material, containing nucleic acids    through the top opening 3 into the basket 7 in the upper sample    space 9 of the at least one container 1;-   (c) Adding enough digestion/lysis buffer {typically a Proteinase K    solution; Commercial examples include PROMEGA DNA IQ Lysis Buffer,    QIAGEN Proteinase K (QIAGEN AG, Garstligweg 8, CH-8634    Hombrechtikon, Switzerland) and INVITROGEN ChargeSwitch® Lysis    Buffer L13 (Invitrogen Corp., 1600 Faraday Avenue, Carlsbad, Calif.    92008, USA)} into the basket 7 so that both the lower fluid space 10    of the container 1 is filled and the upper sample space 9 is also    filled enough to cover the forensic sample 2.    -   The actual volumes will depend on the size of the device—one        imagines different size devices for different sizes of solid        forensics samples 2. In one embodiment, the container 1 would be        of similar dimensions to a typical 2 ml microcentrifuge tube        with a basket that would likely contain a volume of about        1.25 ml. Such an embodiment would likely require adding about 1        to 1.25 ml of buffer in this step.-   (d) Sealing the entire device with a film seal 27, closure 24,    stopper 25, or other means and then allowing to incubate to enable    the extraction process to occur. One would use the incubation times    and temperatures recommended by the manufacturer of the lysis buffer    (temperatures of 56° C., 70° C., and 95° C. are often used for times    ranging up to an hour or more).-   (e) Unsealing or opening the device 1 after incubation and removing    a volume of digestion buffer by accessing the bottom of the lower    fluid space 10 through the essentially vertical channel 11. The    volume removed can be the entire recoverable volume but must be no    less then the volume contained in both the upper sample space 9 plus    the amount of liquid absorbed by the solid forensic sample 2. This    recovered volume of buffer should be set aside in another    vial/tube/etc of the user's choice for later processing.-   (f) Placing the device 1 in a centrifuge and spinning so that any    remaining buffer in the solid forensic sample collects down in the    bottom of the inner chamber 5, preferably in a fluid collecting area    21. The centrifuge should be set at a speed and time that maximizes    recovery of the fluid from the solid forensic sample.-   (g) Removing the remaining volume of digestion buffer that has    collected at the bottom of the lower fluid space 10 by the    centrifugation, preferably in a fluid collecting area 21, by access    through the essentially vertical channel 11. The volume removed is    combined with the volume removed earlier. This sample is now ready    for purification of nucleic acids using commercial nucleic acid    purification kits available from companies such as PROMEGA,    INVITROGEN, QIAGEN, and others.

In this device 1, volumes of lysis buffer containing nucleic acids“washed” off the solid forensic sample 2 are recovered through the“access channel” 11 so that the liquid can be removed and the solidsample 2 left behind. Thus, according to the invention, withdrawal ofthe lysis buffer can be carried out by a pipetting needle withoutrequiring any removal of the basket 7 that contains the solid forensicsample. This particular feature greatly facilitates the utilization ofthe containers 1 in a fully automated workstation such as a TECAN styleworkstation that contains a device for sealing/unsealing andcentrifugation (typical options on these instruments).

Using the containers according to invention, it is optional to either:

-   A Withdraw the extraction liquids and the extracted biological    material with a pipetting needle 12 that is inserted into the    channel 11 and that reaches through the sample space 9 into the    fluid space 10 and to perform amplification and/or analysis of the    extracted biological material outside of the container 1, or to-   B Perform amplification of the extracted biological material in the    fluid space 10 of the container 1.

In any case however, an incubation step preferably is carried out duringthe step (d) and the container 1 is centrifuged after carrying out oneof the step (e).

The biological material to be extracted from the solid forensic samplecan be any sort of biological material. However it is preferred toselect this biological material from a group comprising bodily fluids,cells, DNA, RNA, proteins, microorganisms, and viruses. Particularlypreferred is biological material in the form of nucleic acids ornucleotides.

In order to enable a workstation to carry out the method according tothe invention, such a workstation preferably is equipped with robots andcorresponding control systems as it is known per se in the art of liquidhandling e.g., in pharmaceutical research, where chemical or biochemicalcompounds and/or substances are routinely assayed in regard to potentialpharmaceutical effect.

In such known automatic systems, the transfer and manipulation ofsamples are performed by means of “workstations”, as they are called, orspecial apparatus. These workstations may be operated individually byhand or connected together into an automated system. With automaticsystems, the user does not have to carry out or provide for all theindividual methods of processing. Another common factor uniting suchknown systems lies in the fact that samples are often processed instandardized microplates. Such microplates can be obtained in everypossible format, but typically comprise 96 sample containers or “wells”arranged in a regular 8×12 raster with an interval of 9 mm betweencenters (according to the ANSI standards). Microplates with a multiple,or even only a part, of this number of wells are also used. Differentworkstations may be connected to one or more robots to carry themicroplates. One or more robots, preferably moving in accordance withthe system of Cartesian coordinates, may be used on a workbench top.These robots can carry plates or other sample containers and alsotransfer fluids. A central control system or computer monitors andcontrols these known systems, the outstanding advantage of which lies inthe complete automation of work processes. As a consequence, suchsystems can be operated for hours or days on end, without the need forany human intervention.

Racks for taking up containers 1 in an array preferably have thedimensions of a standard microplate, so that the racks may beautomatically grasped, transported, and placed in a microplate store ora microplate processing station, for example, using a microplatehandling robot. A handling system for providing sample aliquots in rackshaving microplate dimensions is known from the patent EP 0 904 841 B1.The racks used therein have a peripheral frame and a top side and abottom side. These racks comprise latticed partition or intermediatewalls, which define multiple depressions or cavities, each of which iscapable of receiving a sample tube or container. Special retention meansprevent the sample tubes from falling out of their cavities, from whichthey may be removed upward or downward and may also be inserted from thetop or bottom.

Any combinations of the features of the individual embodiments disclosedherein belong to the scope of the present invention. The same referencenumbers are given to the same features shown in the drawings, even whenthey are not specifically addressed in the specification in each case.

List of reference numerals:

-   1 collection/extraction container 23 information tag-   2 solid forensic sample 24 closure-   3 top opening 25 stopper-   4 essentially vertical wall 26 cover-   5 inner chamber 27 film, film structure-   6 bottom 28 collecting set-   7 basket 29 collecting rack-   8 intermediate floor 30 compartments-   9 upper sample space 31 grasping tube-   10 lower fluid space 32 extracting set-   11 essentially vertical channel 33 extracting rack-   12 pipetting needle 34 intermediate wall-   13 top orifice 35 retention means-   14 bottom orifice 36 upper flange-   15 partition wall 37 lower thickened area-   16 upper section of the wall 38 first protrusion-   17 lower section of the wall 39 second protrusion-   18 upper end of lower section 40 off-center axis-   19 socket 41 orientation bar-   20 lower end of upper section 42 orientation grove-   21 fluid collecting area 43 central axis-   22 flat area 44 shoulder

1. A collection/extraction container (1) for collecting solid forensicsamples (2) and/or for extracting biological material from theseforensic samples (2), the container (1) having a top opening (3) for theinsertion of a forensic sample (2), the top opening (3) being surroundedby an essentially vertical wall (4), which encloses an inner chamber (5)that is limited in depth by a bottom (6), the wall (4) and the bottom(6) being impermeable to fluids, wherein the container (1) comprises abasket (7) and an essentially horizontal intermediate floor (8) for theretention of the solid forensic sample (2) during digestion/lysis andextraction, the intermediate floor (8) being permeable to fluids anddividing the inner chamber (5) into an upper sample space (9) and alower fluid space (10), characterized in that the container (1)comprises an essentially vertical channel (11), which is accomplished toallow a pipetting needle (12) to be inserted through the sample space(9) and into the fluid space (10) in that the channel (11) has a toporifice (13) that is situated nearby the top opening (3) of thecontainer (1) and in that the channel (11) penetrates the intermediatefloor (8) in a bottom orifice (14), wherein the container (1) alsocomprises a partition wall (15) that at least partially surrounds thechannel (11) and that separates the latter from the upper sample space(9).
 2. The container (1) of claim 1, characterized in that the wall (4)of the container (1) is essentially cylindrical and in that the channel(11) is arranged off-center and partially formed by the wall (4) and thepartition wall (15) in order to leave as much space as possible for asolid forensic sample (2) that can be introduced into the upper samplespace (9) and that can be kept here by the basket (7).
 3. The container(1) of claim 1, characterized in that the basket (7) of the container(1) is sized to accommodate a buccal swab.
 4. The container (1) of,claim 1 characterized in that the basket (7) is accomplished as a singleinjection molded piece of polymer material and is inserted into theinner chamber (5) of the container (1).
 5. The container (1) of claim 1,characterized in that the basket (7) is accomplished as a part of anupper section (16) of the wall (4) and formed together with this uppersection (16) of the wall (4) as a single injection molded piece ofpolymer material.
 6. The container (1) of claim 1, characterized in thatthe bottom (6) of the container (1) is accomplished to form togetherwith the wall (4) or a lower section (17) of the wall (4) a singleinjection molded piece of polymer material.
 7. The container (1) ofclaim 6, characterized in that the lower section (17) of the wall (4)comprises at its upper end (18) a socket (19) into which a lower end(20) of the upper section (16) of the wall (4) can be sealinglyinserted.
 8. The container (1) of claim 1, characterized in that thebottom (6) and/or the lower section (17) of the wall (4) areaccomplished to form a fluid collecting area (21), which is at a lowerlevel then the rest of the bottom (6) of the container (1), the fluidcollecting area (21) lying on an essentially vertical axis (40) that isdefined by the essentially vertical channel (11) of the container (1).9. The container (1) of claim 1, characterized in that the bottom (6) ofthe container (1) is accomplished as a film structure (27) that is gluedor welded to the wall (4) or to a lower section (17) of the wall (4).10. The container (1) of claim 1 characterized in that the bottom (6)comprises a flat area (22) on its outer side, on or in which flat area(22) an information tag (23) is located, wherein the information tag(23) is selected from a group comprising a 1-D bar code, a 2-D bar code,an RFID transponder, and a RuBee transceiver.
 11. The container (1) ofclaim 1, characterized in that the forensic sample (2) is selected froma group, which comprises a gauze pad or buccal swab, a finding, a filterpaper or WHATMAN FTA paper, and a textile piece.
 12. The container (1)of claim 1, characterized in that it comprises a closure (24) for thetop opening (3), which is selected from a group, which comprises astopper (25), a cover (26), and a film (27).
 13. A collecting set (28)having at least one collection/extraction container (1) of claim 1,characterized in that the collecting set (28) comprises a collectingrack (29) having compartments (30) for receiving containers (1).
 14. Anextracting set (32) having at least one collection/extraction container(1) of claim 1, characterized in that the extracting set (32) comprisesan extracting rack (33) having compartments (30) for receivingcontainers (1).
 15. The collecting set (28) of claim 13, characterizedin that the compartments (30) of this collecting or extracting rack(29,32) are open on the top and bottom, so that the containers (1) areinsertable from above or below into a compartment (30) and are ejectabledownward or upward from this compartment.
 16. The collecting set (28)claim 13, characterized in that the collecting rack (29) has thedimensions of a standard microplate.
 17. A method for extractingbiological material from solid forensic samples (2), the methodcomprising: (a) Providing at least one container (1) with an accessibletop opening (3), an upper sample space (9) with a basket (7), a fluidspace (10), and a channel (11) according to claim 1; (b) Placing solidforensic samples (2) through the top opening (3) into the basket (7) inthe upper sample space (9) of the at least one container; (c) Addingdigestion/lysis buffer into the basket (7) so that both the lower fluidspace (10) of the container (1) is filled and the upper sample space (9)is also filled enough to cover the forensic sample (2); (d) Sealing theentire device (1) and then allowing to incubate to enable the extractionprocess to occur; (e) Unsealing or opening the device (1) afterincubation and removing a volume of digestion buffer by accessing thebottom of the lower fluid space (10) through the essentially verticalchannel (11); (f) Placing the device (1) in a centrifuge and spinning sothat any remaining digestion/lysis buffer in the solid forensic sample(2) collects down in the bottom of the inner chamber (5); (g) Removingthe remaining volume of the digestion/lysis buffer that has collected atthe bottom of the lower fluid space (10) by the centrifugation by accessthrough the essentially vertical channel (11), wherein the solidforensic samples (2) are retained in the basket (7) in the sample space(9) of the at least one container (1) during all the steps (c) through(g).
 18. The method of claim 17, characterized in that the additionalsteps of: (i) Withdrawing the digestion/lysis buffer and the extractedbiological material with a pipetting needle (12) that is inserted intothe channel (11) and that reaches through the sample space (9) into thefluid space (10); and (ii) Performing amplification and/or analysis ofthe extracted biological material outside of the at least one container(1); are carried out.
 19. The method of claim 17, characterized in thatthe digestion/lysis buffer collects down from the solid forensic sample(2) in a fluid collecting area (21) of the bottom of the inner chamber(5).
 20. The method of one of the claims 17, characterized in that thebiological material to be extracted from the solid forensic sample isselected from a group comprising bodily fluids, cells, DNA, RNA,proteins, microorganisms, and viruses.
 21. The method of one of theclaims 17, characterized in that it is performed automatically in aworkstation equipped with robots and corresponding control systems.